G protein-coupled receptor screening assays methods and protocols
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2015
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| Schriftenreihe: | Methods in molecular biology
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| adam_text | Methods in Molecular Biology 1272
John M. Walker, Series Editor
Duarte Miguel F. Prazeres · Sofia Aires M. Martins Editors
G Protein-Coupled Receptor Screening Assays
Methods and Protocols
This volume explores the considerable efforts that have been directed towards the development
of G Protein-Coupled Receptors (GPCR) screening assays in order to disclose GPCR acting com-
pounds, elucidate signaling mechanisms or evaluate compound’s efficacy. New discoveries in the
field, along with the widely recognized need for better and safer pharmaceutical drugs constitute
the main motivation for this book. Readers, both beginners and experienced researchers, will
receive an updated overview of not only the established, but also the innovative technologies
that promise to advance GPCR drug research. This book is organized into two major parts: the
introductory part discusses the necessary foundations for the understanding of GPCR action
and the rationale behind the design of the available screening assays; and part two provides
detailed protocols for different screening approaches. Written in the highly successful Methods
in Molecular Biology series format, the chapters include the kind of detailed description and
implementation advice that is crucial for getting optimal results in the laboratory.
Practical and innovative, G Protein֊ Coupled Receptor Screening Assays: Methods and Protocols
reaches out to everyone involved in the discovery of GPCR-active drugs, and provides a trans-
versal overview of the different levels of GPCR signaling addressable in the different screening
strategies and presents practical examples of how current assay technologies are contributing
to new paradigms in GPCR drug research.
Contents
Preface......................................................................... v
Contributors.................................................................... ix
PartI Introduction
1 G protein-Coupled Receptors: An Overview of Signaling
Mechanisms and Screening Assays................................... 3
Duarte Miguel R Prazeres and Sofia Aires M. Martins
Part II GPCR Screening Assays
2 Time-Resolved FRET Strategy to Screen GPCR Ligand Library......... 23
Nadia Oueslati, Candide Hounsou, Abderazak Belhocine,
Thieric Rodriguez, Elodie Dupuis, Jurriaan M. Zwier, Eric Trinquet
Jean-Philippe Pin, and Thierry Durroux
3 Homogeneous Fluorescence Anisotropy-Based Assay
for Characterization of Ligand Binding Dynamics to GPCRs
in Budded Baculoviruses: The Case of Cv3B-NDP-a-MSH Binding
to MC4 Receptors...................................................... 37
Santa Veiksina, Sergei Kopanchuk, Olga Mazina, ReetLink,
Anne Lille, and Ago Rinken
4 Construction of Recombinant HEK293 Cell Lines
for the Expression of the Neurotensin Receptor NTSR1.................. 51
Su Xiao, Joseph Shiloach, and Reinhard Grisshammer
5 cAMP Assay for GPCR Ligand Characterization:
Application of BacMam Expression System .............................. 65
Olga Mazina, Anni Allibali, Annika Heinloo,
Reet Reinart-Okugbeni, Sergei Kopanchuk, and Ago Rinken
6 Ca2+ Mobilization Assays in GPCR Drug Discovery....................... 79
Grzegorz Woszczek and Elisabeth Fuerst
7 Using Constitutive Activity to Define Appropriate High-Throughput
Screening Assays for Orphan G Protein-Coupled Receptors. ............. 91
Tony Ngo, James LJ. Coleman, and Nicola ƒ. Smith
8 Monitoring G Protein-Coupled Receptor Activation
Using the Protein Fragment Complementation Technique Split TEV........ 107
Michael C Wehry Sabrina Galinski, and Moritz/. Rossner
viii Contents
9 Quantifying GPCR Internalization: A Focus on the Kisspeptin Receptor. 119
Macarena Pampillo and Andy V Babwah
10 GPCR Oligomerization Analysis by Means of BRET and dFRAP............. 133
francisco Ciruela and Víctor Fernández-Dueñas
11 Use of Image J to Recover Information from Individual Cells
in a G Protein-Coupled Receptor Assay................................ 143
Jodo R.C. Trabuco, Sofia Aires M. Martins,
and Duarte Miguel F. Frazer es
12 Methods to Immobilize GPCR on the Surface of SPR Sensors............. 173
Laura Martínez-MuñoZy Rubén Barroso, Anabel Guedán Paredes,
Mario Mellado, and José Miguel Rodríguez-Frade
13 Olfactory Receptor Screening Assay Using NanovesicleTmmobilized
Carbon Nanotube Transistor........................................... 189
Jong Hyun Lim, Juhun Park, Seunghun Hong, and Tat Hyun Park
14 Label-Free Biosensor Assays in GPCR Screening........................ 199
Manuel Grundmann and Evi Kostenis
15 Multidimensional GPCR Profiling and Screening
Using Impedance-Based Label-Free and Real-Time Assay................. 215
Ning Ke, Khanh Nguyen, Jeffery Irelan, and Tama A. Abassi
16 Label-Free Functional Selectivity Assays.............................. 227
Ann M. Ferrie, Vasiliy Goral, Chaoming Wang, and Ye Fang
17 Measurement of Surface-Mediated Ca2+ Transients
on the Single-Cell Level in a M
Contents
Preface......................................................................... v
Contributors.................................................................... ix
PartI Introduction
1 G protein-Coupled Receptors: An Overview of Signaling
Mechanisms and Screening Assays................................... 3
Duarte Miguel R Prazeres and Sofia Aires M. Martins
Part II GPCR Screening Assays
2 Time-Resolved FRET Strategy to Screen GPCR Ligand Library......... 23
Nadia Oueslati, Candide Hounsou, Abderazak Belhocine,
Thieric Rodriguez, Elodie Dupuis, Jurriaan M. Zwier, Eric Trinquet
Jean-Philippe Pin, and Thierry Durroux
3 Homogeneous Fluorescence Anisotropy-Based Assay
for Characterization of Ligand Binding Dynamics to GPCRs
in Budded Baculoviruses: The Case of Cv3B-NDP-a-MSH Binding
to MC4 Receptors...................................................... 37
Santa Veiksina, Sergei Kopanchuk, Olga Mazina, ReetLink,
Anne Lille, and Ago Rinken
4 Construction of Recombinant HEK293 Cell Lines
for the Expression of the Neurotensin Receptor NTSR1.................. 51
Su Xiao, Joseph Shiloach, and Reinhard Grisshammer
5 cAMP Assay for GPCR Ligand Characterization:
Application of BacMam Expression System .............................. 65
Olga Mazina, Anni Allibali, Annika Heinloo,
Reet Reinart-Okugbeni, Sergei Kopanchuk, and Ago Rinken
6 Ca2+ Mobilization Assays in GPCR Drug Discovery....................... 79
Grzegorz Woszczek and Elisabeth Fuerst
7 Using Constitutive Activity to Define Appropriate High-Throughput
Screening Assays for Orphan G Protein-Coupled Receptors. ............. 91
Tony Ngo, James LJ. Coleman, and Nicola ƒ. Smith
8 Monitoring G Protein-Coupled Receptor Activation
Using the Protein Fragment Complementation Technique Split TEV........ 107
Michael C Wehry Sabrina Galinski, and Moritz/. Rossner
viii Contents
9 Quantifying GPCR Internalization: A Focus on the Kisspeptin Receptor. 119
Macarena Pampillo and Andy V Babwah
10 GPCR Oligomerization Analysis by Means of BRET and dFRAP............. 133
francisco Ciruela and Víctor Fernández-Dueñas
11 Use of Image J to Recover Information from Individual Cells
in a G Protein-Coupled Receptor Assay................................ 143
Jodo R.C. Trabuco, Sofia Aires M. Martins,
and Duarte Miguel F. Frazer es
12 Methods to Immobilize GPCR on the Surface of SPR Sensors............. 173
Laura Martínez-MuñoZy Rubén Barroso, Anabel Guedán Paredes,
Mario Mellado, and José Miguel Rodríguez-Frade
13 Olfactory Receptor Screening Assay Using NanovesicleTmmobilized
Carbon Nanotube Transistor........................................... 189
Jong Hyun Lim, Juhun Park, Seunghun Hong, and Tat Hyun Park
14 Label-Free Biosensor Assays in GPCR Screening........................ 199
Manuel Grundmann and Evi Kostenis
15 Multidimensional GPCR Profiling and Screening
Using Impedance-Based Label-Free and Real-Time Assay................. 215
Ning Ke, Khanh Nguyen, Jeffery Irelan, and Tama A. Abassi
16 Label-Free Functional Selectivity Assays.............................. 227
Ann M. Ferrie, Vasiliy Goral, Chaoming Wang, and Ye Fang
17 Measurement of Surface-Mediated Ca2+ Transients
on the Single-Cell Level in a Microfluidic Lab-on֊a-Chip Environment. 247
Michael Kirschbaum, Magnus S. Jaeger, and Claus Duschl
18 Cell-Based Assays and Animal Models for GPCR Drug Screening.......... 257
Hideo Takakura, Mitsuru Hattori, Miho Tanaka, and Takeaki Ozawa
19 Computer-Aided Design of GPCR Ligands................................ 271
Hugo Gutiérrez-de-Terán, Henrik Keränen, Jhonny Azuaje,
David Rodriguez, Johan Àqvist, and Eddy Sotelo
Index
293
|
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| spellingShingle | G protein-coupled receptor screening assays methods and protocols Methods in molecular biology Protéines G / Analyse / Méthodologie ram Protéines / Analyse / Méthodologie ram Récepteurs couplés aux protéines G. Récepteurs couplés aux protéines G / physiologie Récepteurs couplés aux protéines G / analyse Biochemische Analyse (DE-588)4255721-5 gnd G-Protein gekoppelter Rezeptor (DE-588)7723480-7 gnd |
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| title | G protein-coupled receptor screening assays methods and protocols |
| title_auth | G protein-coupled receptor screening assays methods and protocols |
| title_exact_search | G protein-coupled receptor screening assays methods and protocols |
| title_full | G protein-coupled receptor screening assays methods and protocols ed. by Duarte Miguel F. Prazeres ... |
| title_fullStr | G protein-coupled receptor screening assays methods and protocols ed. by Duarte Miguel F. Prazeres ... |
| title_full_unstemmed | G protein-coupled receptor screening assays methods and protocols ed. by Duarte Miguel F. Prazeres ... |
| title_short | G protein-coupled receptor screening assays |
| title_sort | g protein coupled receptor screening assays methods and protocols |
| title_sub | methods and protocols |
| topic | Protéines G / Analyse / Méthodologie ram Protéines / Analyse / Méthodologie ram Récepteurs couplés aux protéines G. Récepteurs couplés aux protéines G / physiologie Récepteurs couplés aux protéines G / analyse Biochemische Analyse (DE-588)4255721-5 gnd G-Protein gekoppelter Rezeptor (DE-588)7723480-7 gnd |
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